Urbanczyk-Wochniak, Ewa , Huhman, David V. , Lei, Zhentian , Dobritsa, Anna , Swanson, Robert , Edlund, Anna F. , Preuss, Daphne , Sumner, Lloyd W. .
Application of anatomically resolved metabolomics to elucidate function of gene involved in Arabidopsis thaliana pollen exine assembly.
Presented work is a part of NSF 2010 project, the main aim of which is to identify the genes required to construct exine, the outer wall of pollen grains. Exine plays important role in early stages of pollen-stigma binding and also is critical for pollen survival. During screening of broad spectrum of mutants for exine deficient phenotypes a set of candidate genes was identified. For testing purposes of candidate genes involved in excine biosynthesis metabolomics approach was applied. Metabolomics has great advantage for gene function elucidation, but most of the published methods focused on bulk tissues, which could sometimes reduce obtained information. Therefore, we decided to collect dissected part of the Arabidopsis flowers (e.g. anthers) from wild-type and three different lap (for less adherent pollen) mutants and analyzed by ultraperformance liquid chromatography/time-of-flight mass spectrometry (UPLC/TOFMS). UPLC/TOFMS method has the substantial benefit of higher separations and resolution, which enables a greater visualization of the metabolome, specifically secondary metabolism part. Extraction and analysis methods were down-sized according to ours previous reports to accommodate smaller quantities of materials such as those to be encountered with dissected floral organs. Currently, we are able to identify approximately 150 different components in antherís extracts collected from 50 Arabidopsis inflorescences. For method validation and new gene function discovery comparative profiling between wild-type and mutants in genes coding chalcone synthases, lap5 , lap6 and double cross lap5_6, were performed. Differential level of several secondary metabolites including phenolics, fatty acids, and yet unidentified compounds is presented and discussed.
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1 - Samuel Roberts Noble Foundation, Plant Biology Division, Ardmore, Oklachoma, 73401
2 - Samuel Roberts Noble Foundation, Plant Biology Division
3 - University of Chicago, The Devision of Biological Sciences
4 - Valparaiso University, Department of Biology
5 - Spelman College, Departament of Biology
Presentation Type: Plant Biology Abstract
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM